Fatty Acids of Lipoproteins CagT and Lpp20

Overview

Journal Microbiol Spectr

Specialty Microbiology

Date 2024 Mar 19

PMID 38501821

Authors

Mark S McClain

William E Boeglin

Holly M Scott Algood

Alan R Brash

Affiliations

Soon will be listed here.

Abstract

Bacterial lipoproteins are post-translationally modified by the addition of acyl chains that anchor the protein to bacterial membranes. This modification includes two ester-linked and one amide-linked acyl chain on lipoproteins from Gram-negative bacteria. lipoproteins have important functions in pathogenesis (including delivering the CagA oncoprotein to mammalian cells) and are recognized by host innate and adaptive immune systems. The number and variety of acyl chains on lipoproteins impact the innate immune response through Toll-like receptor 2. The acyl chains added to lipoproteins are derived from membrane phospholipids. membrane phospholipids have previously been shown to consist primarily of C14:0 and C19:0 cyclopropane-containing acyl chains. However, the acyl composition of lipoproteins has not been determined. In this study, we characterized the acyl composition of two representative lipoproteins, Lpp20 and CagT. Fatty acid methyl esters were prepared from both purified lipoproteins and analyzed by gas chromatography-mass spectrometry. For comparison, we also analyzed phospholipids. Consistent with previous studies, we observed that the phospholipids contain primarily C14:0 and C19:0 cyclopropane-containing fatty acids. In contrast, both the ester-linked and amide-linked fatty acids found in lipoproteins were observed to be almost exclusively C16:0 and C18:0. A discrepancy between the acyl composition of membrane phospholipids and lipoproteins as reported here for has been previously reported in other bacteria including and . We discuss possible mechanisms.IMPORTANCEColonization of the stomach by is an important risk factor in the development of gastric cancer, the third leading cause of cancer-related death worldwide. persists in the stomach despite an immune response against the bacteria. Recognition of lipoproteins by TLR2 contributes to the innate immune response to . However, the role of lipoproteins in bacterial persistence is poorly understood. As the host response to lipoproteins depends on the acyl chain content, defining the acyl composition of lipoproteins is an important step in characterizing how lipoproteins contribute to persistence.

References

Kelly T, Stachula S, Raetz C, Anderson M . The firA gene of Escherichia coli encodes UDP-3-O-(R-3-hydroxymyristoyl)-glucosamine N-acyltransferase. The third step of endotoxin biosynthesis. J Biol Chem. 1993; 268(26):19866-74. View

Folch J, Lees M, SLOANE STANLEY G . A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957; 226(1):497-509. View

Jiang X, Duan Y, Zhou B, Guo Q, Wang H, Hang X . The Cyclopropane Fatty Acid Synthase Mediates Antibiotic Resistance and Gastric Colonization of Helicobacter pylori. J Bacteriol. 2019; 201(20). PMC: 6755722. DOI: 10.1128/JB.00374-19. View

Almagro Armenteros J, Tsirigos K, Sonderby C, Petersen T, Winther O, Brunak S . SignalP 5.0 improves signal peptide predictions using deep neural networks. Nat Biotechnol. 2019; 37(4):420-423. DOI: 10.1038/s41587-019-0036-z. View

Takeuchi H, Islam J, Kaneko A, Kimura A, Shida T, Oboshi W . protein that binds to and activates platelet specifically reacts with sera of -associated chronic immune thrombocytopenia. Platelets. 2021; 32(8):1120-1123. DOI: 10.1080/09537104.2021.1945570. View

Hirai Y, Haque M, Yoshida T, Yokota K, Yasuda T, Oguma K . Unique cholesteryl glucosides in Helicobacter pylori: composition and structural analysis. J Bacteriol. 1995; 177(18):5327-33. PMC: 177327. DOI: 10.1128/jb.177.18.5327-5333.1995. View

Babu M, Priya M, Selvan A, Madera M, Gough J, Aravind L . A database of bacterial lipoproteins (DOLOP) with functional assignments to predicted lipoproteins. J Bacteriol. 2006; 188(8):2761-73. PMC: 1446993. DOI: 10.1128/JB.188.8.2761-2773.2006. View

OToole P, Janzon L, Doig P, Huang J, Kostrzynska M, Trust T . The putative neuraminyllactose-binding hemagglutinin HpaA of Helicobacter pylori CCUG 17874 is a lipoprotein. J Bacteriol. 1995; 177(21):6049-57. PMC: 177441. DOI: 10.1128/jb.177.21.6049-6057.1995. View

Agnihotri G, Crall B, Lewis T, Day T, Balakrishna R, Warshakoon H . Structure-activity relationships in toll-like receptor 2-agonists leading to simplified monoacyl lipopeptides. J Med Chem. 2011; 54(23):8148-60. PMC: 3228886. DOI: 10.1021/jm201071e. View

10.

Bartling C, Raetz C . Steady-state kinetics and mechanism of LpxD, the N-acyltransferase of lipid A biosynthesis. Biochemistry. 2008; 47(19):5290-302. PMC: 2435086. DOI: 10.1021/bi800240r. View

11.

Fischer W, Puls J, Buhrdorf R, Gebert B, Odenbreit S, Haas R . Systematic mutagenesis of the Helicobacter pylori cag pathogenicity island: essential genes for CagA translocation in host cells and induction of interleukin-8. Mol Microbiol. 2002; 42(5):1337-48. DOI: 10.1046/j.1365-2958.2001.02714.x. View

12.

Hanzelmann D, Joo H, Franz-Wachtel M, Hertlein T, Stevanovic S, Macek B . Toll-like receptor 2 activation depends on lipopeptide shedding by bacterial surfactants. Nat Commun. 2016; 7:12304. PMC: 4974576. DOI: 10.1038/ncomms12304. View

13.

Jin M, Kim S, Heo J, Lee M, Kim H, Paik S . Crystal structure of the TLR1-TLR2 heterodimer induced by binding of a tri-acylated lipopeptide. Cell. 2007; 130(6):1071-82. DOI: 10.1016/j.cell.2007.09.008. View

14.

Couture L, Piao W, Ru L, Vogel S, Toshchakov V . Targeting Toll-like receptor (TLR) signaling by Toll/interleukin-1 receptor (TIR) domain-containing adapter protein/MyD88 adapter-like (TIRAP/Mal)-derived decoy peptides. J Biol Chem. 2012; 287(29):24641-8. PMC: 3397892. DOI: 10.1074/jbc.M112.360925. View

15.

Li J, Lee D, Madrenas J . Evolving Bacterial Envelopes and Plasticity of TLR2-Dependent Responses: Basic Research and Translational Opportunities. Front Immunol. 2013; 4:347. PMC: 3808894. DOI: 10.3389/fimmu.2013.00347. View

16.

Wyckoff T, Lin S, Cotter R, Dotson G, Raetz C . Hydrocarbon rulers in UDP-N-acetylglucosamine acyltransferases. J Biol Chem. 1998; 273(49):32369-72. DOI: 10.1074/jbc.273.49.32369. View

17.

Baj J, Korona-Glowniak I, Forma A, Maani A, Sitarz E, Rahnama-Hezavah M . Mechanisms of the Epithelial-Mesenchymal Transition and Tumor Microenvironment in -Induced Gastric Cancer. Cells. 2020; 9(4). PMC: 7225971. DOI: 10.3390/cells9041055. View

18.

Pailler J, Aucher W, Pires M, Buddelmeijer N . Phosphatidylglycerol::prolipoprotein diacylglyceryl transferase (Lgt) of Escherichia coli has seven transmembrane segments, and its essential residues are embedded in the membrane. J Bacteriol. 2012; 194(9):2142-51. PMC: 3347048. DOI: 10.1128/JB.06641-11. View

19.

Bukholm G, Tannaes T, Nedenskov P, Esbensen Y, GRAV H, Hovig T . Colony variation of Helicobacter pylori: pathogenic potential is correlated to cell wall lipid composition. Scand J Gastroenterol. 1997; 32(5):445-54. DOI: 10.3109/00365529709025079. View

20.

Lin A, McClain M, Beckett A, Caston R, Harvey M, Dixon B . Temporal Control of the Helicobacter pylori Cag Type IV Secretion System in a Mongolian Gerbil Model of Gastric Carcinogenesis. mBio. 2020; 11(3). PMC: 7327173. DOI: 10.1128/mBio.01296-20. View